Multicolor super-resolution imaging with photo-switchable fluorescent probes
Multicolor super-resolution imaging with photo-switchable fluorescent probes
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DOI：
10.1126/science.1146598
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发表时间：
2007-09-21
期刊：
影响因子：
56.9
通讯作者：
Zhuang, Xiaowei
中科院分区：
文献类型：
--
作者：
Bates, Mark;Huang, Bo;Zhuang, Xiaowei
在远场光学纳米显微镜的最新进展，使荧光成像的空间分辨率为20至50纳米。然而，多色超分辨率成像仍然是一项具有挑战性的任务。在这里，我们介绍了一个家庭的光开关荧光探针，并展示了随机光学重建显微镜（STORM）。每个探针由可在荧光和暗态之间循环的光可切换的“报告”荧光团和促进报告的光活化的“活化剂”组成。报告子和激活子的组合配对允许创建具有许多不同颜色的探针。这些探针的稀疏子集的迭代，颜色特异性激活允许它们以纳米精度定位，从而能够构建超分辨率STORM图像。使用这种方法，我们展示了DNA模型样品和哺乳动物细胞的20- 30纳米分辨率的双折射成像。这项技术将有助于在纳米尺度上直接观察分子间的相互作用。
Recent advances in far-field optical nanoscopy have enabled fluorescence imaging with a spatial resolution of 20 to 50 nanometers. Multicolor super-resolution imaging, however, remains a challenging task. Here, we introduce a family of photo-switchable fluorescent probes and demonstrate multicolor stochastic optical reconstruction microscopy (STORM). Each probe consists of a photo-switchable "reporter" fluorophore that can be cycled between fluorescent and dark states, and an "activator" that facilitates photo-activation of the reporter. Combinatorial pairing of reporters and activators allows the creation of probes with many distinct colors. Iterative, color-specific activation of sparse subsets of these probes allows their localization with nanometer accuracy, enabling the construction of a super-resolution STORM image. Using this approach, we demonstrate multicolor imaging of DNA model samples and mammalian cells with 20- to 30-nanometer resolution. This technique will facilitate direct visualization of molecular interactions at the nanometer scale.